CN106198996B - Early diagnosis biomarker, kit and its application of minimal change nephrosis - Google Patents
Early diagnosis biomarker, kit and its application of minimal change nephrosis Download PDFInfo
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- CN106198996B CN106198996B CN201610487763.2A CN201610487763A CN106198996B CN 106198996 B CN106198996 B CN 106198996B CN 201610487763 A CN201610487763 A CN 201610487763A CN 106198996 B CN106198996 B CN 106198996B
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/34—Genitourinary disorders
- G01N2800/347—Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy
Abstract
The invention discloses a kind of biomarker STMN1 albumen of early diagnosis of minimal change nephrosis, the kit containing anti-STMN1 protein antibodies and its application.STMN1 high expression in early stage is without the glomerular injury of light microscopic structure change, STMN1 albumen and gene expression amount are also more significantly raised than in unmarred cell in the glomerular podocyte of damage, and STMN1 albumen can be used for the early diagnosis of the minimal change nephrosis without obvious structure change under the microscope.Using the kit of the present invention, after immunohistochemical staining, STMN1 protein positive cells dye brown, can observe to obtain under an optical microscope.This method is simple to operate accurate, to the less demanding of sample, available for extensive examination or retrospective study.
Description
Technical field
The invention belongs to biotechnology and medical domain, and in particular to a kind of life of the early diagnosis of minimal change nephrosis
Substance markers thing, kit and its application.
Background technology
Minimal change nephrosis (minimal change nephropathy, MCD) is with High-grade Proteinuria, hypoproteinemia
Disease, oedema, hyperlipidemia are one of polytype of nephrotic syndrome of essential characteristic.Children are apt to occur in, account for childhood idiopathic
80% or so of nephrotic syndrome, account for the 5%-10% of adult with primary nephrotic syndrome.Minimal change nephrosis is to sugared cortex
The treatment of hormone is sensitive, but easily recurs, and is easily changed into Steroid-Resistance or hormone dependant, clinical treatment is more intractable.And should
Disease emphasizes individualized treatment, i.e., adjusts therapeutic scheme for the pathological change of Different Individual and the same individual different courses of disease.
Therefore, the correctly early diagnosis to minimal change nephrosis is particularly important.Minimal change nephrosis under light microscopic glomerulus without
Obvious lesion, immunofluorescence is negative, is only sexually revised by feature under Electronic Speculum, i.e., extensive glomerulus visceral layer epithelial cell podocytic process
Merge to be made a definite diagnosis.And electron microscopy check-up is costly, diagnosis duration, current domestic medical institutions are not yet general extensively
And.The characteristics of this is sick determines the difficulty of pathological diagnosis, so as to easily miss the best occasion for the treatment of patient.Meanwhile occurring
When Steroid-Resistance or hormone dependant, then the change of histological type should be paid attention in time.Therefore, one kind can be effectively and simple in early stage
The appearance of the method for easy diagnosis minimal change nephrosis, it is most important to patient.
On the other hand, in the different pathological types of renal glomerular disease, rust removability nephrosis (membranous
Nephropathy, MN) and minimal change nephrosis both, the anomalous variation of most glomerulus structures visible under light microscopic.Face
Bed membranous nephropathy can be diagnosed by immunofluorescence technique, but the relatively common microscope of immunofluorescence microscopy is expensive.
If the sick characteristic lesion can be observed under common light microscopic by immunohistochemical staining, not only facilitate
Discriminating to two kinds of diseases, it helps the diagnosis of membranous nephropathy is further quickly determined, so as to be clinicians make pin
Reliable foundation is provided to the therapeutic scheme of property.
The content of the invention
In view of defect present in above-mentioned prior art, it is an object of the invention to provide a kind of minimal change nephrosis
Early lesion has close relationship and can be independently used for detecting the biomarker of minimal change nephrosis early lesion, and will
It is applied to the early diagnosis kit of manufacture minimal change nephrosis.
The present invention seeks to by following technical solution to realize.
A kind of early diagnosis biomarker of minimal change nephrosis, the biomarker are STMN1 albumen.
The present invention also provides described biomarker in reagent of the manufacture for the early diagnosis of minimal change nephrosis
Application in box, it comprises the following steps：
(1) preparation is derived from the testing sample and reference substance of subject；
(2) the STMN1 albumen in the testing sample and reference substance of step (1) is quantitative determined, obtain sample value and
Control value；
(3) sample value and the control group that are obtained in step (2) are compared, the sample value is higher than described
Control value, then judge the subject with minimal change nephrosis.
Further, the testing sample of described subject is derived from the nephridial tissue with glomerular injury patient, described right
The normal kidney tissue of nephrectomy patient is derived from according to product.
Further, quantitative survey is carried out to the STMN1 albumen using immunohistochemistry staining method in step (2)
It is fixed.Staining power and the positive cell of the testing result of immunohistochemistry staining method according to STMN1 protein positive cells
Account in the percentage of all same cells and judge.According to the immunohistochemical staining for STMN1 albumen in the present invention
Qualitative results, it can also judge whether subject is with minimal change nephrosis patient.
Testing sample described above and reference substance can be any kind of in vitro nephridial tissue or cell sample, including new
Renal needle biopsy tissue, the formalin of fresh collection fix the nephridial tissue of FFPE, the freezing nephridial tissue of OCT embeddings, liquid nitrogen
What is preserved is fresh without fixed nephridial tissue etc..No matter which type of biological specimen is applied to, those skilled in the art can root
The pre-treating method of sample is determined according to prior art information, makes the sample after processing be applied to utilize immunohistochemistry staining method
Qualitative and quantitative determination is carried out to the STMN1 albumen.
Kit of the present invention, including anti-STMN1 antibody.As long as anti-STMN1 antibody has special to STMN1 albumen
Associativity, then its species or source etc. are not particularly limited.Furthermore it is possible to it is that polyclonal antibody or monoclonal resist
Body.
Further, the kit also includes the confining liquid and STMN1 antibody specificity knots of closing testing sample tissue
The biomarker secondary antibody of conjunction, the developer with biomarker secondary antibody progress chromogenic reaction.It is direct, easy, highly sensitive in order to detect
The purpose of degree, it is preferential to use the method using the secondary antibodies for being combined with label, combined using secondary antibodies with mark substance
The indirect detection method such as the method for the polymer formed.
Secondary antibodies herein refer to that resisting STMN1 antibody has the antibody of specific binding.Secondary antibodies can use
From the commercially available secondary antibodies with label of rabbit, goat, mouse etc..
Further, the confining liquid contains 5wt% lowlenthal serums, 1wt%BSA, 0.01wt% polyethylene glycol octyl group benzene
The phosphate buffer of base ether and pH7.4, the biomarker secondary antibody are biotinylation secondary antibody, and the developer contains horseradish enzyme mark
Remember strepto- avidin and DAB colour reagents.
Further, the kit also includes the brazilwood extract dyeing liquid as counterstain.
Further, the kit also includes the antigen being made up of 0.01M citrate buffers and phosphate buffer
Repair system, and the elimination endogenous peroxydase system being made up of 3% hydrogen peroxide and phosphate buffer.
In the present invention, the result of study of animal Glomerular lesions is shown, STMN1 is in kidney of the early stage without light microscopic structure change
High expression, is further confirmed, STMN1 albumen and gene expression amount exist through western blotting and qPCR in bead damage
It is also more significantly raised than in unmarred cell in the glomerular podocyte of damage.It is normally and common using China with reference to this result
The patient specimen of three kinds of Glomerular lesions, including MN, MCD, FSGS (FSGS), using simple and easy
Immunohistochemical method, qualitatively confirm the biomarker effect that STMN1 early diagnoses in minimal change nephrosis.At present
Also without (STMN1) albumen of Stathmin 1 and the report of the relevance of minimal change nephrosis.
The present invention carries out immuning tissue using the kit is utilized, for the testing sample and reference substance for being derived from subject
Chemical staining, determine the expression of testing sample and the STMN1 albumen in reference substance.After immunohistochemical staining, STMN1 albumen
Positive cell dyes brown, can observe to obtain under an optical microscope.The disease of nephridial tissue is judged according to sample staining conditions
Diagnosis：The diagnosable brown positive reaction of nucleus and cell body of glomerular podocyte is minimal change nephrosis.This method
It is simple to operate accurate, to the less demanding of sample, available for extensive examination or retrospective study.These methods can apply to
A variety of samples, frozen tissue, paraffin-embedded tissue, Fresh tissue etc., it is possible to provide multi-level detection information, can be small
The research of the glomerular disease such as pathotype nephrosis provides support, also provides dependable detection for clinical diagnosis and direction of medication usage
Index.So that patient obtains more fully, effectively treating, the purpose for improving disease prognosis is finally reached.
Brief description of the drawings
Fig. 1 represents expressions of the STMN1 in Adriamycin-induced Nephropathy renal tissues of rats；
Fig. 2 represents expressions of the STMN1 in minute nephropathy clinical patient nephridial tissue；
Fig. 3 represents expression contrast situations of the STMN1 in the nephridial tissue of different types of glomerular injury Disease.
Embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these examples be merely to illustrate the present invention and
It is not used in the application of the limitation present invention.The test method of unreceipted specific experiment condition in example below, generally according to
Condition described in normal condition, or according to the condition provided on product description.Material used, reagent etc. in example below
Unless otherwise specified, commercially obtain.
1. testing sample and reference substance：
(1) testing sample and reference substance from experimental animal：
According to document (Okuda S. etc., Kidney international, 1986；29(2):Side described in 502-510)
Method, by rat (male Sprague-Dawley rat 6~8 weeks, purchased from from Dalian Medical Univ's Experimental Animal Center), give two
Secondary tail vein injection adriamycin (4.0 mgs/kg and 2.5 mgs/kg), obtains adriamycin-induced nephropathy in rats rat model, takes
Nephridial tissue, as glomerulus nephrosis group (testing sample).The physiological saline that reference substance equally measures from tail vein injection twice
Renal tissues of rats.
(2) testing sample and reference substance from patient：
The nephridial tissue of the patient of common three kinds of Glomerular lesions is derived from as testing sample, common three kinds of glomerulopathies
It is changed into minimal change nephrosis (MCD), membranous nephropathy (MN) and FSGS (FSGS).It is derived from nephrectomy
The normal kidney tissue of patient is as reference substance.
2. using the kit containing anti-STMN1 antibody, exempt from for the testing sample and reference substance for being derived from subject
Epidemic disease histochemical stain, determine the expression of testing sample and the STMN1 albumen in reference substance.After immunohistochemical staining,
STMN1 protein positive cells dye brown, can observe to obtain under an optical microscope.Judge kidney group according to sample staining conditions
The medical diagnosis on disease knitted：The diagnosable brown positive reaction of nucleus and cell body of glomerular podocyte is minute lesion type kidney
Disease.
(1) kit of anti-STMN1 antibody, including the confining liquid of closing testing sample tissue and STMN1 antibody spy are contained
Biomarker secondary antibody and biomarker secondary antibody that the opposite sex combines carry out the developer of chromogenic reaction.Preferentially, the confining liquid contains
There are the phosphate buffer of 5wt% lowlenthal serums, 1wt%BSA, 0.01wt% Triton X-100 and pH7.4, the life
Substance markers secondary antibody is biotinylation secondary antibody, and the developer contains horseradish enzyme mark strepto- avidin and DAB colour reagents.
The kit can also include the brazilwood extract dyeing liquid as counterstain.
The kit can also include the antigen retrieval system being made up of 0.01M citrate buffers and phosphate buffer
System, and the elimination endogenous peroxydase system being made up of 3% hydrogen peroxide and phosphate buffer.The kit can pin
It is applied to property the nephridial tissue sample of FFPE.
3. immunohistochemical staining and criterion
Immunohistochemical staining is carried out to testing sample and reference substance using kit of the present invention, entered according to coloration result
The early diagnosis of row minimal change nephrosis, comprises the following steps：
1. close：In the tissue plus 200 μ l confining liquids of pending sample, it is incubated at room temperature 60 minutes；
2. antigen-antibody reaction：
(1) primary antibody is incubated：Organizationally plus 150 μ l are according to 1:The STMN1 antibody of 100 dilution proportions, is covered with sealed membrane
Tissue, in 4 DEG C of overnight incubations, then with PBS 3 times, every time 10 minutes；
If negative control group, STMN1 antibody is replaced with same volume confining liquid.
(2) secondary antibody is incubated：Organizationally plus 150 μ l biotinylation secondary antibody working solutions, it is incubated at room temperature 60 minutes, PBS 3
It is secondary, 10 minutes every time；
3. horseradish enzyme mark strepto- avidin working solution (S-A/HRP) is organizationally added dropwise, it is incubated at room temperature 60 minutes, PBS
Cleaning 3 times, every time 10 minutes；
4. DAB colour developings (use commercially available DAB dyeing liquors)：
(1) Substrate cocktail is prepared：In the distilled water of 1ml pH value=7.0, a drop reagent A is added, after being well mixed,
Reagent B and reagent C each one is added dropwise to wherein, is well mixed to obtain Substrate cocktail；
(2) develop the color：150 μ l Substrate cocktails are added dropwise organizationally, color development at room temperature 3~10 minutes, fully after colour developing certainly
Water rinses terminating reaction；
5. dyeing, dehydration, transparent, mounting：Histotomy is immersed into brazilwood extract dyeing liquid 2 times, every time 2 seconds；Then with steaming
Distilled water is handled 3 times, every time 5 minutes；Again successively with respectively processing 1 time, every time 1 minute of 50% ethanol, 70% ethanol, 90% ethanol；
Again 2 times are handled with absolute ethyl alcohol, 2 minutes every time, dimethylbenzene and absolute ethyl alcohol 1:1 mixture processing 1 time, 2 minutes, 100% 2
Toluene is handled 2 times, every time 2 minutes；Finally with gummy mounting, preserve；
6. the medical diagnosis on disease of nephridial tissue is judged according to sample staining conditions：The nucleus and cell body of glomerular podocyte be in
Diagnosable brown positive reaction is minimal change nephrosis.
It is applied to appoint on the theoretical method of the early diagnosis for carrying out nephridial tissue minimal change nephrosis of the invention described above
The tumor tissue in vitro or cell sample of what type, including the renal needle biopsy tissue of fresh collection, formalin fix paraffin
Embed nephridial tissue, the freezing nephridial tissue of OCT embeddings, Liquid nitrogen storage it is fresh without fixed nephridial tissue etc..No matter which kind of class is applied to
The biological specimen of type, those skilled in the art can determine the pre-treating method of sample according to prior art information, make place
Sample after reason be applied to it is of the present invention and SABC detection.For several typical samples and its processing method difference
It is：
(1) method of the early diagnosis minimal change nephrosis is applied to formalin fixation FFPE nephridial tissue
When, the pretreatment to sample comprises the following steps：
A. dewax：Formalin fix FFPE nephridial tissue conventional section, the paraffin tissue sections that 4~5 μm of thickness according to
It is secondary to be dewaxed 3 times, every time 5 minutes by 100% dimethylbenzene；Absolute ethyl alcohol dewaxes 3 times, every time 2 minutes；95% ethanol, 90% second
Respectively dewaxing 1 time, 2 minutes of alcohol, 70% ethanol, 50% ethanol；
B. antigen retrieval：Antigen retrieval is carried out with 0.01M citrate buffers, then with PBS 3 times, every time 5 points
Clock；
C. endogenous peroxydase is eliminated：Histotomy is placed in 3% hydrogen peroxide, is incubated at room temperature 30 minutes, is used
PBS 3 times, 5 minutes every time.
Processing gained sample is directly used in detection.
(2) during the nephridial tissue for the freezen protective that the method for the early diagnosis minimal change nephrosis is applied to OCT embeddings,
Pretreatment to sample comprises the following steps：
A. tissue is fixed：By patient's renal needle biopsy tissue about 2x2cm3It is put immediately into 4% paraformaldehyde, 4 DEG C, it is fixed
24 hours；
B.PBS is cleaned 3 times, every time 10 minutes；
C. tissue is immersed into 30% sucrose, 4 DEG C overnight；
D.OCT mixtures (opti-mum cutting temperature compound) embed, and are placed on dry ice；
E. after the solidification of OCT mixtures, 7~10 μm of cryostat frozen section；
(3) method of the early diagnosis minimal change nephrosis is applied to the fresh without fixed nephridial tissue of Liquid nitrogen storage
When, the pretreatment to sample comprises the following steps：
A. the fresh nephridial tissue of patient's renal needle biopsy is inserted in liquid nitrogen rapidly, it is stand-by；
B. the used time first immerses tissue in 4% paraformaldehyde, 4 DEG C, fixes 24 hours；
Any one of the laboratory equipment according to possessed by user, feasible above two method：I.e. FFPE or
After OCT embedding process, processing gained sample is directly used in detection.
The STMN1 of embodiment 1 high expression in early stage is without the glomerular injury of light microscopic structure change
Glomerular injury testing sample of the early stage without light microscopic structure change prepares：
The Adriamycin-induced Nephropathy rat that adriamycin obtains is injected, sacrificed by decapitation, abdominal cavity is splitted, takes out a part of kidney group
Knit, normal saline flushing, and be cut into fritter, after the fixation of the fixers such as formalin, FFPE, Fu Er is prepared
Malin fixes FFPE nephridial tissue, as lesion group.Instead of the rat of adriamycin injecting normal saline, put to death, take out kidney group
Knit, the method same with lesion group processing, obtain formalin and fix FFPE nephridial tissue, as a control group.
The FFPE nephridial tissue of obtained lesion group and control group will be prepared, upper microtome, obtain 5 μ m thicks
Paraffin tissue sections, for hematoxylin eosin staining (HE dyeing) and immunohistochemical staining, see under an optical microscope
Examine coloration result.
Figure 1A is HE coloration results, visible in Figure 1A, and relative to control group, the glomerulus of lesion group is in structure without obvious
Change.
Figure 1B is STMN1 immunohistochemical staining results, and visible in Figure 1B, relative to control group, STMN1 is in lesion group
Glomerular podocyte in high expression, hardly express STMN1 in the sertoli cell in the glomerulus of control group, but in glomerulus
In the model of damage, the high expression STMN1 of most of glomerular podocyte (in figure part STMN1 positive cells arrow tables
Show).
The lesion group and the EPON812 resin embeddings nephridial tissue section of control group that Fig. 1 C obtain for electric Microscopic observation, Fig. 1 C
In it is visible, relative to control group, the Podocytes in Renal Tissue of lesion group, it is seen that projection fusion basal membrane thickening (indicated by an arrow).
On the Research foundation of above-mentioned STMN1 qualitative expressions, in order to further confirm that STMN1 in glomerular injury nephridial tissue
In quantitative expression, by the nephridial tissue of Adriamycin-induced Nephropathy rat and normal kidney tissue (control group), extract total protein, adopt
With the expression of Westen Blotting methods measure STMN1 albumen；Total serum IgE is extracted simultaneously, is detected using quantifying PCR method
The expression of STMN1 genes.Westen Blotting results such as Fig. 1 D, quantitative PCR result such as Fig. 1 E, it is seen that STMN1 egg
White and gene expression amount is more significantly raised than in unmarred cell in the glomerular podocyte of damage.
As a result show：STMN1 can be used as a biomarker, and the glomerular injury of animal imitating examine in early days
It is disconnected.Even if damage is to there is no any sign in conventional light microscopic, only under the Electronic Speculum of the amplification at 800,000 times, the observable qualitative change in side
Change.Summary qualitative and quantitative interpretation of result, STMN1 can be as a biologies for being used for minute nephropathy early diagnosis
Mark.
Expression of the STMN1 of embodiment 2 in minute nephropathy
Fig. 2 represents expression of the STMN1 in minute nephropathy.
Minute nephropathy patient testing sample prepares：The clinical kidney of minute nephropathy patient wears sample, through formal
After the fixers such as woods are fixed, FFPE, formalin is prepared and fixes FFPE nephridial tissue, as lesion group.Kidney is cut
Except the same method processing of the normal kidney tissue of art patient, obtain formalin and fix FFPE nephridial tissue, as a control group.
The FFPE nephridial tissue of obtained lesion group and control group will be prepared, upper microtome, obtain 5 μ m thicks
Paraffin tissue sections, for immunohistochemical staining, coloration result is observed under an optical microscope.
As a result show：Relative to control group, the glomerulus of lesion group changes in om observation in structure without obvious.But
Compared with control group, STMN1 is expressed in the glomerulus of minute nephropathy sample group substantially to be increased, its positive brown expression master
It is distributed in the nucleus and cell body of sertoli cell (part STMN1 positive cells are indicated by an arrow in enlarged drawing).And compare
Glomerular podocyte in group shows that STMN1 radiolucent tables reach.Fig. 2 result is visible, and STMN1 can be used for even in light microscope
Under no any structure change minute nephropathy early diagnosis biomarker.
Fig. 3 represents expression contrast situations of the STMN1 in the nephridial tissue of different types of glomerular injury Disease.
Clinically it is diagnosed as minimal change nephrosis (MCD), membranous nephropathy (MN) and FSGS
(FSGS) the fresh nephridial tissue of patient's renal needle biopsy, as lesion group.The normal kidney tissue of nephrectomy patient, as right
According to group, after inserting the fixation of the fixers such as formalin, FFPE, formalin is prepared and fixes FFPE nephridial tissue,
Upper microtome, obtain the paraffin-embedded tissue section of 5 μM of thickness, the immunohistochemical staining for STMN1.
Immunohistochemical staining result such as Fig. 3 of STMN1 in control group, MCD, MN and FSGS, Fig. 3 results can
See, compared with control group, STMN1 is obvious in minimal change nephrosis (MCD) and the glomerular podocyte of membranous nephropathy (MN)
Enhancing.STMN1 only expresses nucleus and cell body in sertoli cell in the glomerulus tissue of small nephrosis (MCD), but in film
In the glomerulus tissue of property nephrosis, STMN1 is also significantly expressed and is being looped around except the nucleus and cells in vitro in sertoli cell
In sertoli cell projection around blood vessel button loop (part STMN1 positive cells are indicated by an arrow in enlarged drawing).STMN1 is in FSGS
Expression is negative.
In the present invention, STMN1 is expressed in glomerular injury nephridial tissue and significantly improved, can be positive with preliminary judgement, STMN1
The diagnosable nephridial tissue of reaction is glomerular injury disease.Can be further true in the expression and distribution of sertoli cell according to STMN1
Surely it is the glomerular injury of which kind of lesion.STMN1 only expresses the nucleus and cell body in sertoli cell, without obvious elongated protrusion
Diagnosable around the nephridial tissue of blood vessel button loop surrounding is minimal change nephrosis.For elementary test teacher, immunofluorescence can be passed through
Technology excludes membranous nephropathy.MN, MCD and FSGS are clinically most commonly seen glomerular injury diseases.FSGS generally can be
Diagnosed under the light microscopic of HE dyeing, MN need to make a definite diagnosis under the fluorescence microscope of immunofluorescence dyeing, but MCD minute lesion types
The diagnosis of nephrosis can only by Electronic Speculum, inspection fee is high, diagnosis duration and popularization be limited (in addition to advanced teaching hospital,
General hospital does not have Electronic Speculum).Based on this discovery, the application provides a series of sides for being used to early diagnose minimal change nephrosis
Method, and related agent combination.The method of the invention can pass through immunohistochemical staining under simple microscope
Mode, it is quick, sensitive, intuitively minimal change nephrosis is diagnosed and the discriminating with membranous nephropathy and FSGS.This hair
Bright described operating method is simply accurate, to the less demanding of sample, available for extensive examination or retrospective study.These sides
Method can apply to a variety of samples, frozen tissue, paraffin-embedded tissue, Fresh tissue etc., it is possible to provide multi-level detection
Information, support can be provided for the research of the glomerular disease such as minimal change nephrosis, also be carried for clinical diagnosis and direction of medication usage
For dependable Testing index.So that patient obtains more fully, effectively treating, the mesh for improving disease prognosis is finally reached
's.
Claims (2)
1.STMN1 albumen is being manufactured for the application in the kit of the early diagnosis of minimal change nephrosis, it is characterised in that
Comprise the following steps：
(1) preparation is derived from the testing sample and reference substance of subject；
(2) the STMN1 albumen in the testing sample and reference substance of step (1) is quantitative determined, obtains sample value and control
Value；
(3) sample value and the control value that are obtained in step (2) are compared, the sample value is higher than the control
Value, then judge the subject with minimal change nephrosis；
The testing sample of described subject is derived from the nephridial tissue with glomerular injury patient, and the reference substance is derived from the nephrectomy
The normal kidney tissue of art patient.
2. application according to claim 1, it is characterised in that immunohistochemistry staining method pair is used in step (2)
The STMN1 albumen is quantitative determined.
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| CN116558929B (en) * | 2023-07-07 | 2023-09-19 | 中日友好医院(中日友好临床医学研究所) | Podocyte immunofluorescence staining method and application thereof |
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| CN1712542B (en) * | 2004-06-25 | 2012-07-04 | 中国科学院上海生命科学研究院 | Screen and use for labelled proto-protein 18 of protein molecule related to hepatocellular carcinoma |
| PT2666015T (en) * | 2011-01-21 | 2017-03-22 | Basilea Pharmaceutica Ag | Use of stathmin as a biomarker of drug response to furazanobenzimidazoles |
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